Design, Develop, Validate, and Implement Biomarkers for Clinical Investigations

设计、开发、验证和实施用于临床研究的生物标志物

• 批准号: 8158346
• 负责人: Liang Cao
• 金额: $ 19.1万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8158346
• 关键词: Biological Markers; Clinical Trials; design

Background: The development of targeted anti-cancer therapies requires the demonstration of the effects of the investigation agents on the intended target and pathway and the discovery of the markers that can be associated with the response. Thus, we are engaged in detecting the effects of agents on pathway-specific biomarkers and in discovering response biomarkers to be implemented in correlative studies. I. Technology, Assay Design, Development, and Validation. We develop, validate, and implement assays for clinical specimens using electrochemiluminescence (ECL)-based immunoassays. This is the most sensitive and quantitative immunoassay technology platform today. The ECL platform is well suited for this ongoing task because it offers a high degree of flexibility, stability and reliability. It is capable of multiplex analysis to determine the levels of total and phospho-proteins in a single assay well using a limited amount of clinical specimens. Because clinical samples may vary dramatically, the ability to normalize these samples beyond total protein concentration is critical in generating statistically significant data with patient specimens. At the present, we developed, validated and utilized a range of biomarker assays to address the following areas in clinical investigations: 1. Angiogenic factors in plasma and up-regulation of HIF1a in hypoxic tissues for Anti-angegenic agents 2. Cytokines and growth factors for varies targeted agents 3. Cell surface receptors changes as a results of the administration of targeted therapeutic antibodies 4. Intracellular phosphoproteins for a varies kinase inhibitors 5. Apoptotic biomarkers for targeted and cytotoxic agents II. Current and Recently Completed Biomarker Studies. Currently, we are engaged with 10 clinical protocols at NCI-CCR. For many of these clinical trials, we helped to design, develop, validate, and implement customized biomarker assays for correlative analytical studies. The following are some examples: 1) For a phase II clinical trial of sorafenib in androgen-independent prostate cancer (PI, William Dahut), our laboratory performed assay validation with the investigational agent sorafenib and implemented assays with bone marrow biopsy specimens to determine its effects on the targeted MAP kinase pathway (Dahut WL, et al. Clin. Cancer Res. 14: 209-14, 2008). 2) For another clinical trial that investigated a combination targeted therapy against ovarian cancer (PI, Elise Kohn), our group developed assays and performed the analysis of angiogenic factor vascular endothelial growth factor (VEGF) and cytokines, including interleukin-6 (IL6) and interleukin-8 (IL8) from plasma obtained from the patients throughout the drug trial period (Azad NS, et al. J. Clin. Oncol. 26: 3709-14, 2008). 3) In another rapamycin trial (PI, Chand Khanna), we measured the changes in the relative changes of S6RP signaling from both osteosarcoma biopsies and peripheral blood mononuclear cells (PBMC) following the administration of rapamycin. Our data showed a highly statistically significant reduction of p-S6RP in PBMC. The p/t-S6RP ratio changed from 25% pre-drug to 0.5% post-drug with p less than 0.001, with samples obtained from day 2 to the end of the study. Our results further showed that there is a high degree of variation in tumor biopsies. The statistically significant effect of rapamycin on the S6RP pathway can only be demonstrated as p/t-S6RP ratio using our duplex assay capable of quantitatively measuring both p-S6RP and t-S6RP in the same assay well (Paoloni MC et al., PLoS One, 5:e11013, 2010).

背景：有针对性的抗癌疗法的发展需要证明调查剂对预期目标和途径的影响以及可以与反应相关的标记物的发现。因此，我们参与检测药物对途径特异性生物标志物的影响，并发现在相关研究中实施的反应生物标志物。 I.技术，测定设计，开发和验证。我们使用电化学发光（ECL）的免疫测定法开发，验证和实施临床标本的测定法。这是当今最敏感和定量的免疫测定技术平台。 ECL平台非常适合这项正在进行的任务，因为它具有高度的灵活性，稳定性和可靠性。它能够使用有限量的临床标本来确定单个测定中的总磷酸蛋白水平和磷酸蛋白质的水平。由于临床样品可能会发生巨大变化，因此将这些样品归一化的能力超出了总蛋白质浓度，对于用患者标本生成统计学上重要的数据至关重要。 目前，我们开发，验证和利用了一系列生物标志物测定法，以解决临床研究中的以下领域：1。抗盖剂组织中血浆中的血管生成因子和HIF1A的血管生成因子和HIF1A的上调2.抗恒温药物2。细胞因子和生长因子的生长因子的各种靶向体3。挑战者3。 A的磷蛋白各种激酶抑制剂5。靶向和细胞毒性剂II的凋亡生物标志物II。当前和最近完成的生物标志物研究。目前，我们在NCI-CCR上使用了10种临床方案。对于许多临床试验，我们帮助设计，开发，验证和实施定制的生物标志物测定法进行相关分析研究。以下是一些例子：1）对于索拉非尼在雄激素独立的前列腺癌（PI，William Dahut）中的II期临床试验中，我们的实验室对研究剂索拉非尼进行了测定验证，并使用骨髓活检进行了测定，以确定其对靶标MAP KINase Pathway的作用，以确定其对靶标的MAP Kinase Pathway的影响（DAHUT cancer and canci cancion and canci cancy and cancy。 2) For another clinical trial that investigated a combination targeted therapy against ovarian cancer (PI, Elise Kohn), our group developed assays and performed the analysis of angiogenic factor vascular endothelial growth factor (VEGF) and cytokines, including interleukin-6 (IL6) and interleukin-8 (IL8) from plasma obtained from the patients throughout the drug trial period (Azad NS, et al. J. Clin。 3）在另一项雷帕霉素试验（PI，Chand Khanna）中，我们测量了帕帕霉素后，S6RP信号传导的相对变化和外周血单核细胞（PBMC）的相对变化。我们的数据显示，PBMC中P-S6RP的统计学上显着降低。 P/T-S6RP的比率从25％的预药物变为0.5％的药物，P小于0.001，从第2天获得的样品到研究结束。我们的结果进一步表明，肿瘤活检有很高的变化。雷帕霉素对S6RP途径的统计显着作用只能使用我们的双链测定法证明为P/T-S6RP比，能够在同一测定中量化P-S6RP和T-S6RP（Paoloni MC等人）（Paoloni MC等人，PLOS ONE，5：E11013，2010）。